Identification of species-specific DNA in feedstuffs

Due to the menace of transmission of spongiform encephalopathies, feed components intended for ruminant nutrition must be checked for the presence of ruminant-derived materials. A sensitive method for the identification of bovine- and ovine- and also swine- and chicken-specific mitochondrial DNA sequences based on Polymerase Chain Reaction (PCR) has been developed. The specificity of the primers for PCR has been tested using samples of DNA of other vertebrate species, which may also be present in rendering plant products intended for feed manufacture. The method allows the detection in concentrate mixtures of 0.01% of the target species derived material. The identity of a sample containing 0.1% of bovine, ovine, swine, and chicken meat-and-bone meal has further been confirmed by sequencing.     

The Impact of an Abandoned Uranium Mining Area on the Contamination of Agricultural Land in its Surroundings

This paper focuses on the influence of uranium tailing piles on the contamination of their immediate surroundings. The monitoring was carried out on arable land near Pribram in Central Bohemia, Czech Republic. The monitored arable land is located 600 m northeast of the bottom of the piles. In addition, the high uranium (U) content in the topsoil ranged from 40 to 220 ppm. A former portion of it was found in an exchangeable fraction (49%). Both of these facts make this land unsuitable for agricultural use. The sequential extraction discovered a shift in the U content among sample spots situated in and outside the path of the predominant wind direction. In the first group, uranium binds mainly to Fe/Mn oxides, while in the other samples, U is accumulated in the more mobilizable fractions.     

Supercritical fluid extraction of bioavailable amino acids from soils and their liquid chromatographic determination with fluorometric detection

A new procedure with supercritical CO2 modified with 0.5 mL of water and 0.75 mL of 0.1 M HCl in situ and 0.75 mL of water on-line at 15 MPa and 50 degrees C for 45 min was applied for the extraction of bioavailable amino acids from soil samples. Total extraction time was 60 min, but more favorable conditions are even possible for selected groups of amino acids. All analytes were trapped into 20 mL of methanol with satisfactory recovery (94-104%) and determined using high-performance liquid chromatography with fluorometric detection on a Zorbax Eclipse column (4.6 x 75 mm, 3.5 microm) with Na2HPO4 and acetonitrile/methanol/water as a mobile phase. Linear calibration curves were obtained (r > 0.999 except 0.99823 for Ile) with lower limits of detection (S/N = 3) in the range from 1.54 pg (Gly) to 13.5 pg (Cy2) or from 18.6 fmol (Ser) to 64.8 fmol (Lys). Validation and repeatability data are also given. Comparable results were obtained with a robust, commonly used extraction method (0.5 M ammonium acetate, 60 min in shaker, followed by filtration and lyophilization). Limiting values of artificial release of amino acids were also determined for each soil sample to eliminate any false results to ensure that all extracted amino acids originate from soil solution and exchangeable bound positions of soil samples.     

Seasonal patterns of cellulase concentration in desert soil

The concentration of cellulose in plant material greatly affects the decomposition rate of plant-derived litter and hence carbon availability. The disappearance of pure cellulose in soil was studied as a measure of plant decomposition and carbon turnover. Our objective was to understand the effect of various cellulose concentrations and plant material added to soil and collected during different seasons, on cellulase concentrations under laboratory conditions (e.g. constant soil moisture and incubation temperature). The percentage of recovery of the enzyme in the control soil and in samples amended with known amounts of cellulose powder was estimated. Several methods for estimating soil cellulase concentrations/activity are available, most based on the determination of released reducing sugars. The method used in this study is based on the cleavage of a cellulose-azure substrate by cellulase to spectrophotometrically detectable fragments. Our results showed a significant correlation (p<0.05) between cellulose concentration and cellulase levels in soil, which varied along the study period. When pure cellulose was added to the soil, cellulase was detected after 7 days of incubation, whereas when plant material was added to the soil, cellulase was detected after 14 days. The recovery of cellulase from soil was also found to be seasonally dependent. The method of cellulase determination used in this study was simple, safe and rapid. From the results presented in this study, it can be assumed that there are seasonally dependent factors that affect the existence and concentration of cellulase in soils of the arid Negev Desert, in addition to organic matter, water and temperature.     

Water Lettuce Pistia stratiotes L. Response to Lead Toxicity

The effects of one of the most toxic heavy metals, lead (Pb), applied in two different concentrations and combined with chelate application were investigated on the water macrophyte (Pistia stratiotes L.) physiology. The influences were observed by the chlorophyll and free amino acid content determination. Also the lead accumulation in macrophyte biomass was investigated to assess the potential efficiency of this plant for rhizofiltration of highly Pb-polluted water. Na EDTA and Na citrate were used as chelates and Pb(NO₃)₂ as lead supplement. The application of organic chelates simulated conditions of an induced phytoextraction process. Statistical analyses were performed as a one-way ANOVA with a subsequent Tukey HSD test at a level of P < 0.05. Pb contents in both root and leaf tissues gradually increased with increasing Pb concentrations in the nutrient solution. More lead was accumulated in leaves than in roots within all treatments. The total chlorophyll content decreased with increased Pb concentration and with a higher content of chelates. The chelate addition increased the total amino acid content in leaves but decreased the total amino acid content in roots. The addition of lead with chelates decreased the dry biomass weight. However, water macrophyte showed extremely high lead accumulation in biomass in the short term (up to 8 days) and this accumulation potential could be used for relatively fast and effective decrease of high concentration of this risk element in contaminated water or sewage.     

Manganese Biogeochemistry in a Central Czech Republic Catchment

Mn biogeochemistry was studied from 1994 to 2003 in a small forested catchment in the central Czech Republic using the watershed mass balance approach together with measurements of internal stores and fluxes. Mn inputs in bulk deposition were relatively constant during a period of sharply decreasing acidic deposition, suggesting that the Mn source was terrestrial, and not from fossil fuel combustion. Mn inputs in bulk deposition and Mn supplied by weathering each averaged 13 mg m^?2 year^?1 (26 mg m^?2 year^?1 total input), whereas Mn export in streamwater and groundwater averaged 43 mg m^?2 year^?1. Thus an additional Mn source is needed to account for 17 mg m^?2 year^?1. Internal fluxes and pools of Mn were significantly greater than annual inputs and outputs. Throughfall Mn flux was 70 mg m^?2 year^?1, litterfall Mn flux was 103 mg m^?2 year^?1, and Mn net uptake by vegetation was 62 mg m^?2 year^?1. Large pools of labile or potentially labile Mn were present in biomass and surficial soil horizons. Small leakages from these large pools likely supply the additional Mn needed to close the watershed mass balance. This leakage may reflect an adjustment of the ecosystem to recent changes in atmospheric acidity.     

Size of protected areas is the main determinant of species diversity in orchids

Efficient allocation of conservation resources will be achieved only if the priorities for biodiversity conservation – the “hotspots” – are correctly defined. To achieve this we need to pinpoint the main determinants of species diversity. Area, energy available and latitude are thought to be the most important determinants of species richness. Area is clearly the most important, but the relative importance of the other two is uncertain. To test the relative importance of energy available and latitude, data on the species richness of orchids was collected for various countries in the world, the influence of area factored out and the residuals correlated with energy available at these countries and with latitude. This was performed for both total area and that of the protected areas for the 67 countries from five continents, in order to determine which gives a better prediction. We show that – at the large scale considered – area is always very important, latitude is more important than energy available and the size of the protected areas gives a better fit than the total area of the country in most cases. This implies that conservation efforts should be directed to maximizing the size of the protected areas in each country.     

Indoor Aerosol Particle Deposition in an Empty Office

The size- and time-resolved indoor/outdoor aerosol concentration relationships were studied experimentally in an empty office without internal particle sources. Two Scanning Mobility Particle Sizers (SMPS) and an Aerodynamic Particle Sizer (APS) sampled alternately from indoor and outdoor, together covering the size range 3–10,000 nm. The results showed that the indoor aerosol concentration depends mainly on the air exchange and deposition rates, and the outdoor concentration. At higher air exchange rates the indoor aerosol concentration approaches the outdoor one. This was observed for the accumulation mode particles. The size-dependent deposition rates were estimated using a simple aerosol dynamics experiment. It was based on equilibration of the indoor/outdoor concentrations by opening windows and analyzing the concentration decay curves after the windows were closed. For this purpose a simple mass balance model was used. The overall loss rate and the asymptotic value of indoor concentration were found by applying a non-linear least squares method on the time dependence of the indoor concentrations. The air exchange rate was estimated from the overall loss rate and the steady state values of the indoor/outdoor concentration of the accumulation mode particles.     

Identification of Resistance Genes Against Powdery Mildew in Four Accessions of Hordeum Vulgare SSP. Spontaneum

Summary Four newly detected accessions of wild barley (Hordeum vulgare ssp. spontaneum) resistant to powdery mildew caused by Blumeria graminis f. sp. hordei were studied with the aim of finding the number of genes/loci conferring the resistance of individual accessions, the type of inheritance of the genes and their relationships to the Mla locus. F₂ populations after crosses between the winter variety ‘Tiffany’ and four wild barley accessions and use of microsatellite DNA markers were focused on the identification of individual resistance genes/loci by means of their chromosomal locations. In PI466495, one locus conferring powdery mildew resistance was identified in highly significant linkage with the marker Bmac0213. This location is consistent with the known locus Mla on chromosome 1HS. In the other three accessions the resistance was determined by two independent loci. In PI466197, PI466297 and PI466461, one locus was identified on chromosome 1HS and three new loci were revealed on chromosomes 2HS (highly significant linkage with Bmac0134), 7HS (highly significant linkage with Bmag0021) and 7HL (significant linkage with EBmac0755). Our prospective aim is identification of further linked DNA markers and the exact location of the resistance genes on the barley chromosomes.